H5N1亚型禽流感灭活疫苗和DNA疫苗诱导IFN-γ应答评价  

Evaluation of interferon-γresponses in chickens immunized with inactivated vaccine or HA DNA vaccine against H5N1 avian influenza virus

作　　者：戴华 徐正中 陈俊华 季琰 焦新安 

机构地区：[1]扬州大学江苏省人兽共患病学重点实验室/江苏省动物重要疫病与人兽共患病防控协同创新中心/医学院,江苏扬州225009

出　　处：《扬州大学学报（农业与生命科学版）》2017年第3期1-5,10,共6页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家自然科学基金资助项目(30972171;81472815);江苏省优势学科建设工程项目(2014-05)

摘　　要：用H5N1亚型禽流感病毒(AIV)油乳剂灭活疫苗和携带H5N1亚型AIV血凝素(HA)的重组DNA疫苗pVAX1-HA分别免疫2周龄SPF鸡,共免疫3次,每次间隔4周。每次免疫后14d,每组随机取4只鸡,分离其脾脏淋巴细胞,用H5N1亚型AIV JS株灭活全病毒作为特异性体外刺激抗原刺激鸡脾脏淋巴细胞,72h后,检测其培养上清中特异性ChIFN-γ的水平,同时用HI试验检测免疫鸡血清抗体。结果表明:重组DNA疫苗pVAX1-HA免疫组在二次免疫后,可检测出特异性ChIFN-γ的分泌和一定水平的抗体应答;而H5N1亚型AIV灭活疫苗免疫组均未能检测出ChIFN-γ,二次免疫后能产生较高水平的抗体应答。说明DNA疫苗可激发较高水平的IFN-γ应答,而AIV灭活疫苗主要激发抗体应答,提示两者联合免疫可使免疫的禽群诱导产生强的细胞免疫应答和抗体应答,为AIV的防控提供了参考。Two-week-old SPF chickens were immunized with inactivated vaccine of HbN1 subtype avian influenza virus （AIV） and recombinant plasmid pVAX1-HA containing hemagglutinin （HA） cDNA of H5N1 subtype of AIV （H5N1- AIV）, respectively. Chickens were immunized three times at a 4 weeks＇ interval. At the 14th day post each immuniza- tion, 4 chickens per group were picked randomly, their splenocytes were collected and stimulated using heat-inactivated whole AIV JS strain as specific H5N1-AIV recall antigens. Subsequently, ChIFN-γ response of each immunized chicken was detected by a ChlFN-γ sandwish ELISA after 72 h stimulation of the recall antigens. The hemagglutination inhibition （HI） titers of serum antibodies were measured. Results showed that DNA vaccine pVAX1-HA could be capable of elici- ring specific ChlFN-γ response to H5N1 AIV recall antigens after the second immunization and protective antibodies can he detected with low HI titers. While the chickens vaccinated with H5N1-AIV inactivated vaccine displayed strong anti- body responses with high HI titer especially after the second immunization, but no detectable ChlFN-y after each immuni- zation. Our study indicated that DNA vaccine was good at eliciting cell-mediated immunity represented by the production of IFN-γ while inactivated vaccine mostly produces strong humoral immune responses, and with the superiority of inacti- vated vaccine combined with DNA vaccine strategy, the immunized poultry could elicit not only high cell-mediated immu- nity but also strong antibody response. These results provided a basis for prevention and control of AIV.

关 键 词：H5N1亚型禽流感病毒 灭活疫苗 DNA疫苗 ChIFN-γ 抗体 

分 类 号：S855.3[农业科学—临床兽医学]